Brain-chip modeling neurodegeneration and neuroinflammation in parkinson's disease

Abstract

The invention relates to modeling brain neuronal disease in a microfluidic device, comprising a co-culture of iPS-derived brain endothelial cells; iPS-derived dopaminergic neurons; primary microglia; and primary astrocytes, a Blood-Brain-Barrier (BBB)-Chip and a Brain-Chip. In particular, cross-talk between glial cells (e.g. microglia and astrocytes) with neuronal cells, in further contact with endothelial cells is contemplated for use for identifying drug targets under conditions for inducing in vivo relevant neuronal inflammation, neurodegeneration and neuronal death. Thus, in one embodiment, a microfluidic Brain-Chip comprising a co-culture of brain cells is exposed to α-synuclein preformed fibrils (PFF), a type of pathogenic form of α-synuclein. Such α-synuclein PFF exposure demonstrates an in vivo relevant disease pathogenesis on a microfluidic device as a concentration- and time-controlled manner that may be used for preclinical drug evaluation for diseases related to neuronal inflammation, e.g. Parkinson's disease (PD). In some embodiments, modulation of complement in the presence of neuronal inflammation is contemplated. In some embodiments, drug delivery to brain cells across the BBB is contemplated for preclinical testing of drug efficacy for slowing or stopping neuronal inflammation and degeneration.

1 - 135 . (canceled) 
     
     
         136 . A method, comprising,
 a) providing,   i) an inflammation inducing compound;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels;   iii) a plurality of cells comprising microglial cells mixed with cells, said cells selected from the group consisting of pericytes, astrocytes, and neurons and combinations thereof; and   iv) a population of endothelial cells;   b) culturing said plurality of cells in said first channel and culturing said population of endothelial cells in said second channel;   c) contacting said cultured cells with said inflammation inducing compound under conditions such that cytokine secretion is induced; and   d) detecting said induced cytokine secretion. wherein the amount of cytokine produced is larger than the amount produced in the absence of said microglial cells.   
     
     
         137 . The method of  claim 136 , wherein said inflammation inducing compound is TNF-alpha. 
     
     
         138 . The method of  claim 136 , wherein said cytokine is IL-6. 
     
     
         139 . A method, comprising,
 a) providing,   i) an inflammation inducing compound;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels;   iii) a plurality of cells comprising microglial cells mixed with cells, said cells selected from the group consisting of pericytes, astrocytes, and neurons and combinations thereof; and   iv) a population of endothelial cells;   b) culturing said plurality of cells in said first channel and culturing said population of endothelial cells in said second channel; and   c) contacting said cultured cells with said inflammation inducing compound under conditions such that inflammation is induced, wherein said contacting comprises flowing said inflammation inducing compound into said second channel; and   d) detecting transcriptomic changes in the cells.   
     
     
         140 . The method of  claim 139 , wherein said detecting of transcriptomic changes comprises detecting differentially expressed genes. 
     
     
         141 . The method of  claim 140 , wherein the differentially expressed genes are different from those expressed when said inflammation inducing compound is introduced into said first channel. 
     
     
         142 . The method of  claim 140 , wherein increased expression in a plurality of glia-associated genes is detected only after exposure to the inflammation inducing compound through the second channel. 
     
     
         143 . The method of  claim 142 , wherein said glia-associated genes are selected from the group consisting of GFAP, XYLT1, H19, RGS4, TREM2, PADI2, and PADI4. 
     
     
         144 . The method of  claim 139 , wherein said inflammation inducing compound is TNF-α. 
     
     
         145 . A method, comprising,
 a) providing,   i) α-synuclein (αSyn) fibrils;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels; and   iii) a plurality of dopaminergic neurons in said first channel, said first channel further comprising astrocytes and microglial cells;   b) contacting said dopaminergic neurons with said α-synuclein fibrils; and   c) detecting induced cytokine secretion.   
     
     
         146 . The method of  claim 145 , wherein said contacting comprises flowing said α-synuclein fibrils into said first channel. 
     
     
         147 . The method of  claim 145 , further comprising detecting accumulation of phosphorylated αSyn in said neurons. 
     
     
         148 . The method of  claim 145 , further comprising detecting mitochondrial damage in said neurons. 
     
     
         149 . The method of  claim 145 , further comprising detecting an increase in reactive oxygen species over time. 
     
     
         150 . The method of  claim 145 , further comprising detecting an increase in caspase 3-positive neurons over time. 
     
     
         151 . The method of  claim 145 , further comprising detecting neuroinflammation. 
     
     
         152 . The method of  claim 145 , further comprising detecting apoptosis. 
     
     
         153 . The method of  claim 145 , further comprising detecting neuronal death. 
     
     
         154 . The method of  claim 145 , further comprising detecting microglia activation. 
     
     
         155 . The method of  claim 145 , further comprising detecting astrocyte activation. 
     
     
         156 . The method of  claim 145 , further comprising detecting astrogliosis. 
     
     
         157 . A method, comprising,
 a) providing,   i) α-synuclein (αSyn) fibrils;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels;   iii) a plurality of dopaminergic neurons in said first channel; and   iv) a population of endothelial cells in said second channel;   b) introducing said α-synuclein fibrils into said device; and   c) detecting an inflammatory response of said endothelial cells.   
     
     
         158 . The method of  claim 157 , wherein said αSyn fibrils are introduced into said second channel. 
     
     
         159 . The method of  claim 157 , further comprising detecting accumulation of αSyn in said endothelial cells. 
     
     
         160 . A method, comprising,
 a) providing,   i) α-synuclein (αSyn) fibrils;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels;   iii) a plurality of dopaminergic neurons in said first channel; and   iv) a population of endothelial cells in said second channel;   b) culturing said cells such that said endothelial cells form tight junctions, said tight junctions defining a barrier having a level of permeability;   c) introducing said α-synuclein fibrils into said device; and   d) detecting a change in said level of permeability.   
     
     
         161 . The method of  claim 160 , further comprising c) contacting said endothelial cells with a test compound. 
     
     
         162 . The method of  claim 161 , detecting the impact of said test compound on said permeability. 
     
     
         163 . A method, comprising,
 a) providing,   i) α-synuclein (αSyn) fibrils;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels; and   iii) a plurality of dopaminergic neurons in said first channel;   b) contacting said dopaminergic neurons said α-synuclein fibrils; and   c) detecting neuroinflammation.   
     
     
         164 . The method of  claim 163 , further comprising introducing a drug for testing efficacy for slowing or stopping said neuronal inflammation. 
     
     
         165 . A method, comprising,
 a) providing,   i) an inflammation inducing compound;   ii) a microfluidic device comprising a membrane, said membrane separating first and second microfluidic channels;   iii) a plurality of cells comprising microglial cells mixed with cells, said cells selected from the group consisting of pericytes, astrocytes, and neurons and combinations thereof; and   iv) a population of endothelial cells;   b) culturing said plurality of cells in said first channel and culturing said population of endothelial cells in said second channel;   c) contacting said cultured cells with said inflammation inducing compound under conditions such that inflammation is induced; and   d) introducing a test compound.   
     
     
         166 . The method of  claim 165 , further comprising using biomarkers to determine the effect of said test compound. 
     
     
         167 . The method of  claim 166 , wherein said using of biomarkers comprises gene expression profiling. 
     
     
         168 . The method of  claim 165 , wherein said inflammation inducing compound is TNF-alpha. 
     
     
         169 . The method of  claim 165 , wherein said inflammation inducing compound comprises α-synuclein (αSyn) fibrils.